US 12,270,044 B2
CRISPR systems with engineered dual guide nucleic acids
Ryan T. Gill, Denver, CO (US); Tanya Warnecke, Boulder, CO (US); Andrea Barghetti, Copenhagen (DK); and Line Dahl Poulsen, Ballerup (DK)
Assigned to Celyntra Therapeutics SA, Mont-Saint-Guibert (BE); and DANMARKS TEKNISKE UNIVERSITET, KGS Lyngby (DK)
Filed by Celyntra Therapeutics SA, Mont-Saint-Guibert (BE); and DANMARKS TEKNISKE UNIVERSITET, KGS Lyngby (DK)
Filed on Mar. 24, 2023, as Appl. No. 18/190,063.
Application 18/190,063 is a continuation of application No. 17/963,889, filed on Oct. 11, 2022.
Application 17/963,889 is a continuation of application No. 17/506,572, filed on Oct. 20, 2021.
Application 17/506,572 is a continuation of application No. PCT/US2020/054050, filed on Oct. 2, 2020.
Claims priority of provisional application 62/910,055, filed on Oct. 3, 2019.
Prior Publication US 2023/0407342 A1, Dec. 21, 2023
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 9/22 (2006.01); C12N 15/11 (2006.01); C12N 15/70 (2006.01); C12N 15/85 (2006.01); C12N 15/90 (2006.01)
CPC C12N 15/907 (2013.01) [C12N 9/22 (2013.01); C12N 15/11 (2013.01); C12N 15/70 (2013.01); C12N 15/85 (2013.01); C12N 2310/20 (2017.05); C12N 2800/22 (2013.01)] 27 Claims
 
1. An engineered, non-naturally occurring system comprising:
(a) a targeter nucleic acid comprising, from 5′ to 3′:
(i) a targeter stem sequence;
(ii) a spacer sequence designed to hybridize with a target nucleotide sequence; and
(iii) an optional additional nucleotide sequence;
(b) a modulator nucleic acid comprising a modulator stem sequence complementary to the targeter stem sequence,
wherein the targeter nucleic acid and the modulator nucleic acid are separate nucleic acids, wherein the targeter nucleic acid, the modulator nucleic acid, or both, comprises a chemically modified RNA, and wherein a complex comprising the targeter nucleic acid and the modulator nucleic acid is capable of activating a CRISPR Associated (Cas) nuclease that, in a naturally occurring system, is activated by a single crRNA in the absence of a tracrRNA, wherein the Cas nuclease comprises a Type V-A Cas nuclease; and
(c) the Type V-A Cas nuclease.